多通道流动电泳技术及其在蛋白质、酶和抗体纯化中的应用研究

Multichannel Flow Electrophoresis and Its Applications in Purification of Proteins,Enzymes and Antibodies

提出一种新型制备电泳方法即多通道流动电动电泳技术,建立了微机控制的电泳设备,考查了操作条件对设备分离效率的影响。应用该技术进行牛血清清蛋白（BSA）-牛血红蛋白（HBB）混合物的连续分离,每小时从含BSA、HBB各37.0mg的蛋白质混合物中分离出13.6mg的BSA和20.0mg的HBB。应用该技术进行尿激酶粗品的纯化,可将其比活力由4000IU/mg提高到4×104IU/mg以上,产量在10×104IU/h以上。将该技术与（NH4）SO4沉淀方法结合,从15ml免疫鼠血清中分离出12.5mg高纯度尿激酶抗体IgG,产量为2.1mg/h。上述结果证明多通道流动电泳技术具有分离速度快、精度高和可操作性好等优点,在生物产品分离领域中具有广阔的应用前景。

A new preparative electrophoresis technique,multichannel flow electrophore-sis (MFE) was proposed. A multicompartment electrolyzer and an operation control system used in MFE were developed. The effects of electric field strength, buffering pH and sample introducing flow rate on MFE separation efficiency were examined. The continuous separation of BSA and HBB mixture yielded about 40mg product. The application of MFE was first exemplified by continuous purification of urokinase from urine extracts. 10x104IU/h urokinase product whose specific activity was higher than 40 000IU/mg was obtained per hour. MFE was also applied in the continuous purification of ariti urokinase mouse IgG from mouse serum, which yielded 2. Img mouse IgG hourly with high purity demonstrated by SDS-PAGE. The preliminary results presented in this paper have confirmed the workability of MFE and pointed out its high application potential in biochemical engineering processes.